Biomaterial Database

Protein synthesis in rabbit reticulocytes. Characteristics of mRNA (AUG codon)-dependent binding of Met-tRNAfMet to 40 S and 80 S ribosomes. 1976

B Chatterjee, and A Dasgupta, and S Palmieri, and N K Gupta

The characteristics of Met-tRNAfMet binding to ribosomes (40 S and 80 S) were studied using a two-stage assay method (Gupta, N.K., Chatterjee, B. and Majumdar, A. (1975) Biochem. Biophys. Res. Commun. 65, 797) and the complexes formed were analyzed either by Millipore filtration or by sucrose density gradient centrifugation. The results are summarized as follows: (a) with both assay methods, Met-tRNAfMet binding to 40 S ribosomes was entirely dependent upon addition of a partially purified mixture of initiation factors and AUG codon; (b) this binding occurred over a wide Mg2+ concentration range; significant binding was observed even at 20 mM Mg2+; (c) upon addition of 60 S ribosomes, a significant part of Met-tRNAfMet bound to 40 S ribosomes was transferred to 80 S complex. This transfer reaction had a sharp Mg2+ optimum around 2 mM. Met-tRNAfMet-80 S-AUG complex thus formed was active in Met-puromycin synthesis; (d) Met-tRNAfMet deacylase present in crude 0.5 M KCl ribosomal wash is a potent inhibitor of the binding reaction as it deacylates Met-tRNAfMet in the Met-tRNAfMet-40 S-AUG complex; (e) glutaraldehyde (0.5%) degrades Met-tRNAfMet-40 S-AUG complex but increases the background binding of Met-tRNAfMet to 40 S ribosomes in the absence of AUG codon; (f) polynucleotides containing uracil and adenosine are strong inhibitors of Met-tRNAfMet binding to 40 S ribosomes. The order of inhibitory activities of the polynucleotides tested was as follows: poly(rU)-poly(rA) (2:1) greater than poly(rU)-poly(rA) (1:1) greater than poly(rU) greater than poly(rA). Other RNAs tested such as poly(rC), poly(rI)-poly(rC) and phi6 bacteriophage RNA (double-stranded) were without significant effects on the Met-tRNAfMet-binding reaction.

UI	MeSH Term	Description	Entries
D007700	Kinetics	The rate dynamics in chemical or physical systems.
D008274	Magnesium	A metallic element that has the atomic symbol Mg, atomic number 12, and atomic weight 24.31. It is important for the activity of many enzymes, especially those involved in OXIDATIVE PHOSPHORYLATION.
D008715	Methionine	A sulfur-containing essential L-amino acid that is important in many body functions.	L-Methionine,Liquimeth,Methionine, L-Isomer,Pedameth,L-Isomer Methionine,Methionine, L Isomer
D010442	Peptide Chain Initiation, Translational	A process of GENETIC TRANSLATION whereby the formation of a peptide chain is started. It includes assembly of the RIBOSOME components, the MESSENGER RNA coding for the polypeptide to be made, INITIATOR TRNA, and PEPTIDE INITIATION FACTORS; and placement of the first amino acid in the peptide chain. The details and components of this process are unique for prokaryotic protein biosynthesis and eukaryotic protein biosynthesis.	Chain Initiation, Peptide, Translational,Protein Biosynthesis Initiation,Protein Chain Initiation, Translational,Protein Translation Initiation,Translation Initiation, Genetic,Translation Initiation, Protein,Translational Initiation, Protein,Translational Peptide Chain Initiation,Biosynthesis Initiation, Protein,Genetic Translation Initiation,Initiation, Genetic Translation,Initiation, Protein Biosynthesis,Initiation, Protein Translation,Initiation, Protein Translational,Protein Translational Initiation
D010448	Peptide Initiation Factors	Protein factors uniquely required during the initiation phase of protein synthesis in GENETIC TRANSLATION.	Initiation Factors,Initiation Factor,Factors, Peptide Initiation,Initiation Factors, Peptide
D011817	Rabbits	A burrowing plant-eating mammal with hind limbs that are longer than its fore limbs. It belongs to the family Leporidae of the order Lagomorpha, and in contrast to hares, possesses 22 instead of 24 pairs of chromosomes.	Belgian Hare,New Zealand Rabbit,New Zealand Rabbits,New Zealand White Rabbit,Rabbit,Rabbit, Domestic,Chinchilla Rabbits,NZW Rabbits,New Zealand White Rabbits,Oryctolagus cuniculus,Chinchilla Rabbit,Domestic Rabbit,Domestic Rabbits,Hare, Belgian,NZW Rabbit,Rabbit, Chinchilla,Rabbit, NZW,Rabbit, New Zealand,Rabbits, Chinchilla,Rabbits, Domestic,Rabbits, NZW,Rabbits, New Zealand,Zealand Rabbit, New,Zealand Rabbits, New,cuniculus, Oryctolagus
D012156	Reticulocytes	Immature ERYTHROCYTES. In humans, these are ERYTHROID CELLS that have just undergone extrusion of their CELL NUCLEUS. They still contain some organelles that gradually decrease in number as the cells mature. RIBOSOMES are last to disappear. Certain staining techniques cause components of the ribosomes to precipitate into characteristic "reticulum" (not the same as the ENDOPLASMIC RETICULUM), hence the name reticulocytes.	Reticulocyte
D003062	Codon	A set of three nucleotides in a protein coding sequence that specifies individual amino acids or a termination signal (CODON, TERMINATOR). Most codons are universal, but some organisms do not produce the transfer RNAs (RNA, TRANSFER) complementary to all codons. These codons are referred to as unassigned codons (CODONS, NONSENSE).	Codon, Sense,Sense Codon,Codons,Codons, Sense,Sense Codons
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D001665	Binding Sites	The parts of a macromolecule that directly participate in its specific combination with another molecule.	Combining Site,Binding Site,Combining Sites,Site, Binding,Site, Combining,Sites, Binding,Sites, Combining